caa a22 4500 445853727 CHVBK 20180317145421.0 cr unu---uuuuu 170323e20110401xx s 000 0 eng 10.1007/s00204-010-0587-8 doi (NATIONALLICENCE)springer-10.1007/s00204-010-0587-8 In vitro study of P-glycoprotein induction as an antidotal pathway to prevent cytotoxicity in Caco-2 cells [Elektronische Daten] [Renata Silva, Helena Carmo, Ricardo Dinis-Oliveira, Anabela Cordeiro-da-Silva, Sofia Lima, Félix Carvalho, Maria de Lourdes Bastos, Fernando Remião] The Caco-2 cell line is a reliable in vitro model for predicting drug intestinal absorption and P-glycoprotein (P-gp)-mediated excretion in humans. Recent in vivo studies suggested the induction of P-gp as a cellular protection tool against paraquat poisoning, through the increase in its pulmonary and intestinal excretion. Thus, the aim of the present work was to evaluate P-gp expression and activity in Caco-2 cells exposed to doxorubicin (a known P-gp inducer) and to correlate these changes with paraquat toxic effects. Cytotoxicity of doxorubicin (0-100μM) and paraquat (0-1,000μM) was evaluated for a maximum period of 96h. In doxorubicin-exposed cells, P-gp expression and transport activity were evaluated by flow cytometry, using a fluorescein isothiocyanate-conjugated antibody and the P-gp fluorescent subtract rhodamine 123, respectively. A significant increase in P-gp expression was observed as soon as 6h after exposure to 5μM doxorubicin. P-gp activity also increased after 6h, but only at higher doxorubicin concentrations (over 50μM). Paraquat (0-5,000μM) cytotoxicity was then evaluated with or without previous exposure of the cells to doxorubicin (5-100μM, a concentration range causing both an increase in P-gp expression and activity). Under P-gp induction, a significant reduction in paraquat cytotoxicity was observed. Furthermore, when these cells were incubated with a specific P-gp inhibitor (UIC2 antibody) the doxorubicin protective effects were blocked, confirming the involvement of P-gp in the reduction in paraquat cytotoxicity. In conclusion, the human Caco-2 cell line model can be used for the study of P-gp induction as an antidotal pathway against substrates of this transporter system. Springer-Verlag, 2010 P-glycoprotein induction nationallicence P-glycoprotein transport activity nationallicence Paraquat toxicity nationallicence Caco-2 cells nationallicence BCRP : Breast cancer-resistant protein nationallicence DMEM : Dulbecco's modified Eagle's medium nationallicence DOX : Doxorubicin nationallicence EDTA : Ethylenediamine tetraacetic acid nationallicence FBS : Fetal bovine serum nationallicence FITC : Fluorescein isothiocyanate nationallicence MTT : (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide nationallicence NEAA : Nonessential aminoacids nationallicence NSAIDs : Nonsteroidal anti-inflammatory drugs nationallicence PBS : Phosphate-buffered saline solution nationallicence P-gp : P-glycoprotein nationallicence PQ : Paraquat nationallicence RHO 123 : Rhodamine 123 nationallicence Silva Renata REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut Carmo Helena REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut Dinis-Oliveira Ricardo REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut Cordeiro-da-Silva Anabela Laboratory of Biochemistry, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut Lima Sofia IBMC—Institute of Molecular and Cellular Biology, University of Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal aut Carvalho Félix REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut de Lourdes Bastos Maria REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut Remião Fernando REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut Archives of Toxicology Springer-Verlag 85/4(2011-04-01), 315-326 0340-5761 85:4<315 2011 85 204 https://doi.org/10.1007/s00204-010-0587-8 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00204-010-0587-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Silva Renata REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut NATIONALLICENCE 700 1- Carmo Helena REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut NATIONALLICENCE 700 1- Dinis-Oliveira Ricardo REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut NATIONALLICENCE 700 1- Cordeiro-da-Silva Anabela Laboratory of Biochemistry, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut NATIONALLICENCE 700 1- Lima Sofia IBMC—Institute of Molecular and Cellular Biology, University of Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal aut NATIONALLICENCE 700 1- Carvalho Félix REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut NATIONALLICENCE 700 1- de Lourdes Bastos Maria REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut NATIONALLICENCE 700 1- Remião Fernando REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal aut NATIONALLICENCE 773 0- Archives of Toxicology Springer-Verlag 85/4(2011-04-01), 315-326 0340-5761 85:4<315 2011 85 204 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer